Yamamoto T, Nagae H, Kajihara Y, Terada I
Sea Water Science Research Laboratory, Japan Tobacco Inc., Kanagawa, Japan.
Biosci Biotechnol Biochem. 1998 Feb;62(2):210-4. doi: 10.1271/bbb.62.210.
To supply alpha 2,6-sialyltransferase for the large-scale synthesis of sialoside, we investigated culture conditions for the production of sialyltransferase 0160. The addition of galactose and beef extract, and control of the pH of the culture medium were effective on the production of sialyltransferase 0160. The maximal enzyme productivity reached 550 units/L. Using a crude extract of Photobacterium damsela JT0160 cells as an enzyme source, enzymatic syntheses were performed with mono- and di-saccharides as the sialyl acceptors. It was clarified that a crude extract of P. damsela JT0160 cells can be used as an synthetic catalyst for the enzymatic synthesis of sialyloligosaccharides. Furthermore, the enzyme assay showed that sialyltransferase 0160 could transfer NeuAc to not only N-linked but also O-linked carbohydrate chains. These results indicated that an abundant supply of sialyltransferase 0160 and its broad specificity make possible the synthesis of sialoside on a large scale.
为了提供用于大规模合成唾液酸苷的α2,6-唾液酸转移酶，我们研究了唾液酸转移酶0160的培养条件。添加半乳糖和牛肉提取物以及控制培养基的pH值对唾液酸转移酶0160的产生有效。最大酶产量达到550单位/升。使用美人鱼发光杆菌JT0160细胞的粗提取物作为酶源，以单糖和二糖作为唾液酸受体进行酶促合成。结果表明，美人鱼发光杆菌JT0160细胞的粗提取物可用作酶促合成唾液酸寡糖的合成催化剂。此外，酶活性测定表明，唾液酸转移酶0160不仅可以将NeuAc转移到N-连接的碳水化合物链上，还可以转移到O-连接的碳水化合物链上。这些结果表明，大量供应唾液酸转移酶0160及其广泛的特异性使得大规模合成唾液酸苷成为可能。
